Efficacy of branched-chain hydroxy acids in regulating intestinal microorganisms

Abstract

The present invention relates to the efficacy of branched-chain hydroxy acids in regulating intestinal microorganisms. In the present invention, two branched-chain hydroxy acids were confirmed to exhibit the effect of regulating the growth of intestinal microflora, and were confirmed to promote the growth of probiotic strains, thereby exhibiting a probiotic effect, and to suppress the growth of pathogens. It was also confirmed that two branched-chain hydroxy acids were produced in intestinal microflora, and each branched-chain hydroxy acid was correlated with the intestinal microflora regulation effect.

Description

The intestinal microbiota-regulating effects of branched-chain hydroxy acids

[0001] The present invention relates to the intestinal microflora regulating effect of branched-chain hydroxy acids.

[0002]

[0003] Prebiotics, probiotics, and postbiotics are relatively new terms used to describe a variety of substances that confer health and nutritional benefits to animals. Typically, the term prebiotic refers to substances that stimulate the growth or activity of bacteria in the digestive tract of animals, leading to beneficial health effects. Prebiotics can be selectively fermented ingredients that induce specific changes in both the composition and activity of the gastrointestinal microflora, conferring health benefits to the host. Probiotics generally refer to microorganisms that contribute to the intestinal microbial balance and, consequently, to maintaining health. Many species of lactic acid bacteria (LAB), such as Lactobacillus and Bifidobacterium, are commonly considered probiotics, although some species of Bacillus and some yeasts have also been identified as suitable candidates. Postbiotics refer to non-viable bacterial products or metabolic by-products derived from probiotic organisms that possess biological activity in the host.

[0004] The use of probiotics to improve animal health and nutrition has been shown to be effective in a variety of diseases and health conditions. Furthermore, prebiotics and postbiotics offer potential alternative or adjunctive therapies to the use of live microorganisms. There is growing understanding of the role of prebiotics, probiotics, and postbiotics in modulating immune responses, specifically in regulating the expression of cytokines that control inflammatory responses at both local and systemic levels. For example, ingestion of probiotic bacteria can potentially stabilize the immunological barrier in the gut mucosa by reducing the production of local proinflammatory cytokines. Furthermore, alterations in the characteristics of the indigenous microbiota by probiotic therapy have been shown to reverse some immunological disturbances in human conditions such as Crohn's disease, food allergies, and atopic eczema.

[0005] The human gut microbiome, composed of various phyla, contributes to the metabolic health of several organisms. Various phyla, including Firmicutes, Proteobacteria, Bacteroidetes, and Actinomycetota, dominate the human gut microbiome, and maintaining the balance of gut microbiota has recently been recognized as important for maintaining health. An imbalance in the gut microbiome, known as dysbiosis, is known to cause various metabolic diseases such as obesity, type 2 diabetes, non-alcoholic fatty liver disease, and cardiometabolic diseases, and research on bacterial control is being conducted to restore the balance of the gut microbiome.

[0006] Improving the gut environment provides health benefits for the host, and improving the gut microbiome can be achieved by promoting the growth of probiotics through prebiotics and inhibiting the growth of pathogens through antibiotics. Various metabolites, such as dietary fiber and polyphenols, have been proposed as prebiotics, and recent studies have reported that microbial metabolites can modulate the gut microbiome. For example, short-chain fatty acids produced by gut microbes are known to improve the host gut environment, influence the gut-brain axis, and alter the composition of the gut microbiome. Given the importance of increased probiotic abundance for host health, understanding how these metabolites regulate the growth of the gut microbiome is crucial.

[0007] Accordingly, the inventors of the present invention confirmed that branched-chain hydroxy acids, which are probiotic metabolites, regulate intestinal flora, thereby completing the present invention.

[0008]

[0009] The purpose of the present invention is to provide a food composition for regulating intestinal microflora, which contains branched chain hydroxy acids (BCHA) as an effective ingredient.

[0010] Another object of the present invention is to provide a prebiotic composition comprising branched chain hydroxy acids (BCHA) as an active ingredient.

[0011]

[0012] To achieve the above purpose, the present invention provides a food composition for regulating intestinal microflora, which contains branched chain hydroxy acids (BCHA) as an effective ingredient.

[0013] In addition, the present invention provides a prebiotic composition comprising branched chain hydroxy acids (BCHA) as an active ingredient.

[0014]

[0015] The present invention confirmed that two types of branched-chain hydroxy acids have growth-regulating effects on intestinal microflora, and confirmed that they promote growth in probiotic strains, exerting prebiotic effects while inhibiting the growth of pathogens. Furthermore, it was confirmed that two types of branched-chain hydroxy acids are also produced in intestinal microflora, and the correlation between each type of branched-chain hydroxy acid and its intestinal microflora-regulating effects was confirmed, suggesting that the present invention can be usefully utilized in related industries.

[0016]

[0017] Figures 1 and 2 quantify the proliferation of intestinal flora according to treatment with the branched chain hydroxy acid of the present invention (Figure 1: quantification of HIVA results, Figure 2: quantification of HICA results).

[0018] Figure 3 shows the quantitative analysis of the inter-strain proliferation of intestinal microflora according to treatment with the branched-chain hydroxy acid of the present invention.

[0019] Figures 4 to 7 quantify the growth of beneficial and harmful bacteria according to treatment with the branched chain hydroxy acid of the present invention.

[0020] Figure 4: Confirmation of Lactobacillus plantarum growth

[0021] Figure 5: Confirmation of Lactobacillus fermentum growth

[0022] Figure 6: Confirmation of Lactobacillus salivarius growth

[0023] Figure 7: Confirmation of Bacteroides fragilis growth

[0024] Figures 8 and 9 are diagrams quantifying the production of branched-chain hydroxy acids according to the proliferation of the intestinal flora of the present invention (Figure 8: quantification of HIVA production, Figure 9: quantification of HICA production).

[0025] Figures 10 and 11 are diagrams confirming the correlation between strain growth and the concentration of the branched chain hydroxy acid of the present invention (Figure 10: HIVA correlation analysis, Figure 11: HICA correlation analysis).

[0026]

[0027] Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings. In the following description, detailed descriptions of well-known technologies to those skilled in the art may be omitted. Furthermore, in describing the present invention, detailed descriptions of related known functions or configurations may be omitted if it is determined that such detailed descriptions may unnecessarily obscure the gist of the present invention. Furthermore, the terminology used in this specification is intended to appropriately express preferred embodiments of the present invention, and may vary depending on the intentions of the user or operator, or the customs of the field to which the present invention pertains.

[0028] Therefore, definitions of these terms should be based on the overall content of this specification. Throughout this specification, whenever a part is said to "include" a component, this does not exclude other components, but rather implies the inclusion of additional components, unless otherwise specifically stated.

[0029] The present invention provides a food composition for regulating intestinal microflora, which comprises branched chain hydroxy acids (BCHA) as an effective ingredient.

[0030] The food composition of the present invention may contain, in addition to containing the effective ingredient of the present invention, various flavoring agents or natural carbohydrates as additional ingredients, like conventional food compositions.

[0031] Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and common sugars, and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As the above-mentioned flavoring agent, natural flavoring agent (thaumatin), stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agent (saccharin, aspartame, etc.) can be advantageously used. The food composition of the present invention can be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes, and health supplements.

[0032] In addition, the food composition may contain, in addition to the extract as an active ingredient, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the food composition of the present invention may contain fruit pulp for producing natural fruit juice, fruit juice drinks, and vegetable drinks.

[0033] The functional food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of regulating intestinal microflora. The term "health functional food composition" in the present invention refers to a food manufactured and processed using raw materials or ingredients having functionality useful to the human body according to Act No. 6727 on Health Functional Foods, and means to be consumed for the purpose of obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological effects. The health functional food of the present invention may include conventional food additives, and whether it is suitable as a food additive is determined by the specifications and standards for the relevant item according to the general provisions and general test methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety, unless otherwise specified. Items listed in the "Food Additives Codex" include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Examples thereof include natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high-molecular weight pigment, and guar gum; mixed preparations such as sodium L-glutamate preparations, noodle additive alkaline agents, preservative preparations, and tar color preparations. For example, a health functional food in tablet form can be prepared by mixing the active ingredient of the present invention with excipients, binders, disintegrants, and other additives, granulating the mixture using a conventional method, and then adding a lubricant, etc. to compress and molding, or directly compressing and molding the mixture. In addition, the health functional food in tablet form can contain a maturing agent, etc., if necessary. Among health functional foods in capsule form, hard capsules can be prepared by filling a mixture of the active ingredient of the present invention with additives such as excipients into a conventional hard capsule, and soft capsules can be prepared by filling a mixture of the active ingredient of the present invention with additives such as excipients into a capsule base such as gelatin. The above soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative, etc., as needed.The ring-shaped health functional food can be prepared by molding a mixture of the active ingredient of the present invention with excipients, binders, disintegrants, etc. using a conventionally known method, and, if necessary, can be coated with white sugar or other coating agents, or the surface can be coated with a substance such as starch or talc. The granular health functional food can be manufactured into a granular form using a mixture of the active ingredient of the present invention with excipients, binders, disintegrants, etc. using a conventionally known method, and, if necessary, can contain a flavoring agent, a flavoring agent, etc.

[0034] According to one embodiment of the present invention, the branched chain hydroxy acid may be a compound represented by the following chemical formula 1 or chemical formula 2.

[0035] [Chemical Formula 1]

[0036]

[0037] [Chemical Formula 2]

[0038]

[0039] The compound of the above chemical formula 1 of the present invention may be named 2-hydroxyisovaleric acid and may be a compound with CAS number 17407-56-6.

[0040] The compound of the above chemical formula 2 of the present invention may be named 2-hydroxyisocaproic acid and may be a compound with CAS number 498-36-2.

[0041] According to one embodiment of the present invention, the branched chain hydroxy acid may increase the growth of a strain of a family selected from the group consisting of the Lactobacillaceae family, the Enterococcaccae family, the Lachnosporaceae family, and the Bifidobacteriaceae family.

[0042] According to one embodiment of the present invention, the Lactobacillus strain may be a strain selected from the group consisting of Lactobacillus salivarius, L. plantarum, L. fermentum, L. rhamnosus, and L. sakei.

[0043] According to one embodiment of the present invention, the Enterococcus strain may be Enterococcus faecalis.

[0044] According to one embodiment of the present invention, the Bifidobacterium aceae strain may be Bifidobacterium adolescentis, Bifidobacterium longum, or Bifidobacterium breve.

[0045] According to one embodiment of the present invention, the Lactosporaceae strain may be Agathobacter rectalis or Holdemanella bioformis.

[0046] According to one embodiment of the present invention, the branched chain hydroxy acid may regulate the growth of a strain of the Clostridiaceae family, the Enterobacteriaceae family, or the Bacteroidaceae family.

[0047] According to one embodiment of the present invention, controlling the growth of the Clostridium spp. strain may be inhibiting the growth of Clostridium asparagiforme or Clostridium bolteae, or increasing the growth of Clostridium scindens or Clostridium symbiosum.

[0048] According to one embodiment of the present invention, controlling the growth of the Enterobacteriaceae strain may be inhibiting the growth of Escherichia coli, Salmonella typhimurium, or Providencia stuartii.

[0049] According to one embodiment of the present invention, controlling the growth of the Bacteroidetes strain may be to increase the growth of Bacteroides ovatus or B. uniformis, and to decrease the growth of B. fragilis, B. uniformis, or Phocaeicola dorei.

[0050] In addition, the present invention provides a prebiotic composition comprising branched chain hydroxy acids (BCHA) as an active ingredient.

[0051] The term "prebiotics" as used herein refers to nutrients that promote the growth of beneficial intestinal bacteria, or probiotics. Furthermore, prebiotics can be defined as indigestible food ingredients that promote the growth and activity of beneficial bacteria that contribute to the health of the host. Prebiotics are not digested in the small intestine and migrate to the large intestine, where they can selectively enhance the growth and activity of bacteria, such as lactic acid bacteria, thereby promoting human health. In the large intestine, prebiotics are fermented by intestinal flora, thereby promoting the growth and activity of intestinal flora, which can significantly improve the health of the host.

[0052] Additionally, the present invention provides a method for regulating intestinal microflora, comprising administering to a subject a composition comprising branched chain hydroxy acids (BCHA).

[0053]

[0054] The advantages and features of the present invention, and the methods for achieving them, will become clearer with reference to the detailed examples described below. The present invention will now be described in detail through examples. However, these examples are intended to specifically illustrate the present invention and are not intended to limit the scope of the present invention.

[0055]

[0056] [Experimental Example 1] Preparation for Confirming the Intestinal Bacteria Regulation of Branched-Chain Hydroxy Acids

[0057] 1-1. Reagents used

[0058] High-performance liquid chromatography (HPLC) grade methanol and water were purchased from Thermo Fisher Scientific (USA). All analytical reagents used in the present invention were purchased from Sigma Chemical Co. (USA). Branched-chain hydroxy acids, 2-hydroxyisovaleric acid (HIVA; Chemical Formula 1), 2-hydroxyisocaproic acid (HICA; Chemical Formula 2), N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), methoxyamine hydrochloride, and pyridine were purchased from Sigma (USA). Brain Heart Infusion (BHI) medium was purchased from Kisan Biotech (Korea).

[0059] [Chemical Formula 1]

[0060]

[0061] [Chemical Formula 2]

[0062]

[0063]

[0064] 1-2. Culture conditions of intestinal microbial strains

[0065] The intestinal microbial strains of the present invention are shown in Fig. 3. Specifically, 24 different microbial strains were selected from the Actinobacteria, Firmicutes, Bacteroidetes, and Proteobacteria phyla, which are major phyla present in the human gastrointestinal tract. All strains were then anaerobically cultured on BHI agar medium supplemented with 10% defibrinated horse blood. In addition, the strains cultured on the agar were cultured under anaerobic conditions at 37°C for 24 hours in BHI medium supplemented with 0.05% cysteine. Under anaerobic culture conditions, the strains were cultured in an anaerobic chamber under conditions of 10% CO2, 5% H2, and 85% N2.

[0066]

[0067] 1-3. In vitro screening for the regulation of gut microbiota by branched-chain hydroxy acids.

[0068] The intestinal microbiota cultured for 24 hours under anaerobic conditions at 37℃ was inoculated into a 96-well plate at an OD600 value of 0.01 and treated with BHI medium containing branched-chain hydroxy acids at a concentration of 0.25 mg / ml. The 96-well plate was then sealed to maintain anaerobic conditions, and after incubation at 37℃, the OD value was automatically measured every 15 minutes using a microplate reader. Each value was then normalized and entered into the Baranyi model using Microsoft Excel to present the growth curve for each strain.

[0069]

[0070] 1-4. Confirmation of the prebiotic effect and inhibition of harmful bacteria of branched-chain hydroxy acids.

[0071] Lactiplantibacillus plantarum KACC 11451, Limsilactobacillus fermentum KCTC 15072BP, Ligilactobacillus salivarius KGMB 02057, and Bacteroides fragilis KCTC 5013 were cultured in BHI liquid medium supplemented with 0.05% cysteine ​​at 37°C for 24 hours. Afterwards, each culture was inoculated into a 96-well plate at a concentration of OD600 0.01 and treated with 0.2 ml of BHI medium containing two types of branched-chain hydroxy acids (0.125 to 4.0 mg / ml) or inulin (0.25 mg / ml) as a positive control. Thereafter, the growth of each strain was confirmed using the same method as in Experimental Example 1-3.

[0072]

[0073] 1-5. Quantification of branched-chain hydroxy acids derived from intestinal microorganisms

[0074] Enteric bacteria were cultured in BHI medium supplemented with 0.05% cysteine ​​(5 ml) at 37°C for 24 h. The cultured strains were then inoculated into 96-well plates at an OD600 of 0.01, and 1.5 ml of fresh BHI medium was added. The plates were then cultured in an anaerobic chamber (H25%, CO210%, and N285%) at 37°C for 24 h. After incubation, each culture was collected, centrifuged at 10,000 X g for 10 min, and filtered using a Millex GP 0.22-μm filter (Merck Millipore, Billerica, MA, USA). The filtered supernatant was added to 800 μl of 100% methanol (MeOH), suspended for 30 s, and centrifuged at 4°C and 24,249 X g for 10 min. Afterwards, the supernatant was dried using a high-speed vacuum concentrator, and the dried sample was dissolved in 1 ml of 100% MeOH. To prepare standard solutions, HICA and HIVA stock solutions were serially diluted in 100% MeOH to concentrations of 0.004, 0.008, 0.016, 0.03, 0.06, 0.12, 0.25, 0.5, 0.1, and 0.2 μg / ml, and 2-Chlorophenylalanine was used as an internal standard at a concentration of 10 μg / ml.

[0075]

[0076] 1-6. Gas chromatography-time-of-flight mass spectrometry

[0077] Before analyzing the dried samples in Experimental Examples 1-5 by gas chromatography-time-of-flight-mass spectrometry (GC-TOF-MS), two derivatization steps were performed. 50 μl of methoxyamine hydrochloride dissolved in 20 mg / ml pyridine was added to the dried samples, and the samples were incubated at 30°C and 300 rpm for 90 minutes to perform oxidation. 50 μl of MSTFA was then added to the reaction mixture to induce silylation, followed by incubation at 37°C and 300 rpm for 30 minutes. All samples were analyzed using an Agilent 7890A GC system equipped with an Agilent 7693 autosampler and a Pegasus BT TOF-MS, and were filtered using a Millex GP 0.22-μm filter prior to GC-TOF-MS analysis. The column used for the analysis was an Rtx-5MS capillary column (30 m Х 0.25 mm Х 0.25 μm, Restek Corp., Bellefonte, PA), and a total of 1 μl of the derivatized sample was injected into the GC-TOF-MS with a split ratio of 30:1 for analysis.

[0078]

[0079] 1-7. Statistical Analysis

[0080] Peak areas of branched-chain hydroxy acids were calculated based on the GC-TOF-MS dataset, and their fold-change abundances were represented using heat maps. Significant differences in bacterial growth and production of HIVA and HICA were analyzed using one-way ANOVA with Duncan's multiple range test and independent-samples t-test using PASW Statistics 18 (SPSS Inc., USA). Pearson correlation coefficients between production and growth control of HIVA and HICA were calculated using PASW Statistics 18.

[0081]

[0082] [Example 1] Growth-regulating effects of branched-chain hydroxy acids on intestinal microflora

[0083] To determine the effects of branched-chain hydroxy acids on the intestinal environment, a 96-well plate-based in vitro screening assay was used to determine the regulatory effects of branched-chain hydroxy acids on intestinal microbial growth. Each intestinal strain was cultured as a single cell in liquid medium treated with 0.25 mg / ml of HIVA and HICA, and the ODmax at the end of culture was compared with that of the negative control.

[0084] HIVA significantly increased the growth of Lactobacillaceae and Bifidobacteriaceae, and significantly inhibited the growth of Clostridiaceae and Enterobacteriaceae (Fig. 1).

[0085] HICA showed a tendency to increase the growth of Lactobacillaceae and Bifidobacteriaceae (Fig. 2).

[0086] Clostridiaceae and Bacteroidaceae showed different growth patterns among species when treated with HIVA or HICA, and even within the same family, the growth regulatory effects differed depending on the species. Among the Lactobacillaceae and Bifidobacteriaceae families, L. salivarius and B. adolescentis were the species most stimulated for growth by HIVA or HICA treatment. In the Bacteroidaceae family, the growth of B. ovatus was promoted, and the growth of B. fragilis was inhibited by HICA compared to HIVA. In addition, some strains showed different growth regulatory effects depending on treatment with HIVA or HICA, and the growth of S. typhimurium and P. stuartii was inhibited by HIVA treatment, but did not change by HICA treatment. B. fragilis showed a greater growth reduction in HICA treatment compared to HIVA treatment, and in B. uniformis, HIVA inhibited growth, whereas HICA promoted growth (Fig. 3).

[0087]

[0088] [Example 2] Confirmation of the growth regulation of branched-chain hydroxy acids on probiotics and pathogens.

[0089] To verify the prebiotic and antimicrobial effects of branched-chain hydroxy acids, three probiotic Lactobacillaceae strains, including L. plantarum KACC 11451, L. fermentum KCTC 15027BP, L. salivarius KGMB 02057, and B. fragilis KCTC 5013, and one pathogenic strain, were treated with HIVA or HICA at various concentrations (Fig. 3). As a result, compared with the other strains, L. plantarum was growth-stimulated by HIVA and HICA treatment, and both HIVA and HICA promoted the growth of L. plantarum at the treatment concentrations. Compared with the positive control group, HIVA and HICA showed higher growth promotion effects at 2.0 to 4.0 mg / ml and 0.125 to 2.0 mg / ml, respectively. HIVA showed a significant promotion effect at a concentration of 2.0 to 4.0 mg / ml, and HICA showed a significant effect in the concentration range of 0.25 to 2.0 mg / ml, and in particular, the highest prebiotic effect was confirmed at a concentration of 2.0 mg / ml (Fig. 4).

[0090] In the case of L. fermentum, growth was stimulated when the concentrations of HIVA and HICA were in the ranges of 0.125–1.0 and 0.125–2.0 mg / ml, respectively. Compared with the positive control, HICA showed a higher growth-promoting effect at 0.25–2.0 mg / ml, and HIVA showed a significant effect in the concentration ranges of 0.25–0.5 mg / ml, whereas HICA showed a significant effect in the concentration ranges of 0.125 and 0.5–2.0 mg / ml, but growth was significantly inhibited at a concentration of 2.0–4.0 mg / ml for HIVA and 4.0 mg / ml for HICA (Fig. 5).

[0091] Growth of L. salivarius was stimulated by HIVA and HICA in the concentration ranges of 0.125–4.0 mg / ml and 0.125–2.0 mg / ml, respectively, and showed higher growth than the positive control at concentrations of both metabolites in the ranges of 0.25–2.0 and 0.125–1.0 mg / ml (Fig. 6).

[0092] On the other hand, the pathogenic strain B. fragilis showed no significant difference in growth in the concentration ranges of HIVA and HICA of 0.125 to 1.0 mg / ml and 0.125 to 0.25 mg / ml, respectively, but the growth of B. fragilis significantly decreased as the concentrations of HIVA and HICA increased to 2.0 to 4.0 mg / ml and 0.5 to 4.0 mg / ml, respectively (Fig. 7).

[0093]

[0094] [Example 3] Quantitative production of branched-chain hydroxy acids by intestinal microflora

[0095] This study assessed whether the gut microbiota produces branched-chain hydroxy acids. Specifically, each bacterial species was cultured in BHI medium supplemented with cysteine, and extracellularly secreted branched-chain hydroxy acids were analyzed using GC-TOF-MS.

[0096] As a result, HIVA and HICA were detected in all intestinal microbiota groups, and the production amount showed differences depending on the family of each microbiota (Figs. 8 and 9). The production levels of HIVA and HICA were both significantly higher in the Lactobacillaceae and Lachnosporaceae families, while the production levels in the Clostridiaceae, Enterobacteriaceae, and Bacteroidaceae families were significantly lower compared to the other families. In addition, the Enterococcaceae family showed high HICA production, but the production of HIVA was low, and the Bifidobacteriaceae family showed significantly high HICA production.

[0097]

[0098] [Example 4] Confirmation of the correlation with the growth-regulating effect of branched-chain hydroxy acids.

[0099] To investigate the effects of branched-chain hydroxy acids on the intestinal environment, we analyzed the correlation between their growth-regulating effects on intestinal microorganisms and the production of HIVA and HICA. As shown in Figures 10 and 11, the correlation coefficient was higher for HIVA than for HICA, with HIVA exhibiting a significantly strong positive correlation (r=0.638, p=0.001), and HICA exhibiting a moderate positive correlation (r=0.638, p=0.001).

[0100]

[0101] Therefore, the present invention confirmed that two types of branched-chain hydroxy acids have growth-regulating effects depending on the intestinal microflora, and confirmed that they promote growth in probiotic strains, thus having a prebiotic effect, while inhibiting the growth of pathogens. In addition, it was confirmed that two types of branched-chain hydroxy acids are produced in the intestinal microflora, and the correlation between each type of branched-chain hydroxy acid and the intestinal microflora-regulating effects was confirmed.

Claims

1. A food composition for regulating intestinal microflora containing branched chain hydroxy acids (BCHA) as an active ingredient.

2. A food composition according to claim 1, wherein the branched chain hydroxy acid is a compound represented by the following chemical formula 1 or chemical formula 2. [Chemical Formula 1] [Chemical Formula 2] 3. A food composition according to claim 1, wherein the branched chain hydroxy acid increases the growth of a strain of a family selected from the group consisting of Lactobacillaceae, Enterococcaccae, Lachnosporaceae, and Bifidobacteriaceae.

4. A food composition according to claim 3, wherein the Lactobacillus strain is a strain selected from the group consisting of Lactobacillus salivarius, L. plantarum, L. fermentum, L. rhamnosus, and L. sakei.

5. A food composition according to claim 3, wherein the Enterococcus faecalis strain is Enterococcus faecalis.

6. A food composition according to claim 3, wherein the Bifidobacterium aceae strain is Bifidobacterium adolescentis, Bifidobacterium longum, or Bifidobacterium breve.

7. A food composition according to claim 3, wherein the Lactosporaceae strain is Agathobacter rectalis or Holdemanella bioformis.

8. A food composition according to claim 1, wherein the branched chain hydroxy acid regulates the growth of a strain of the Clostridiaceae family, the Enterobacteriaceae family, or the Bacteroidaceae family.

9. A food composition according to claim 8, wherein the growth of the Clostridium aceae strain is controlled by inhibiting the growth of Clostridium asparagiforme or Clostridium bolteae.

10. A food composition according to claim 8, wherein controlling the growth of the Clostridium spp. strain increases the growth of Clostridium scindens or Clostridium symbiosum.

11. A food composition according to claim 8, wherein the growth of the Enterobacteriaceae strain is controlled by inhibiting the growth of Escherichia coli, Salmonella typhimurium, or Providencia stuartii.

12. A food composition according to claim 8, wherein controlling the growth of the Bacteroidetes strain increases the growth of Bacteroides ovatus or B. uniformis.

13. A food composition according to claim 8, wherein the growth of the Bacteroidetes strain is controlled by reducing the growth of B. fragilis, B. uniformis or Phocaeicola dorei.

14. A prebiotic composition comprising branched chain hydroxy acids (BCHA) as an active ingredient.

15. A method for regulating intestinal microflora, comprising administering to a subject a composition comprising branched chain hydroxy acids (BCHA).

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